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Related Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...

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Recombineering Homologous Recombination Constructs in Drosophila
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An efficient method for recombineering GAL4 and QF drivers.

R Steven Stowers1

  • 1Department of Cell Biology and Neuroscience, Montana State University, Bozeman, MT, USA. sstowers@montana.edu

Fly
|August 23, 2011
PubMed
Summary

New recombineering cassettes enable precise control of gene expression in neurons. This technique facilitates neural circuit mapping by allowing specific targeting of pre- and post-synaptic cells for advanced research.

Area of Science:

  • Neuroscience
  • Molecular Biology
  • Genetics

Background:

  • Independent control of gene expression in pre- and post-synaptic neurons is crucial for neural circuit mapping and manipulation.
  • The GAL4/UAS and Q binary transcription systems offer potential for this precise control.
  • Development of specific GAL4 and QF drivers for neurotransmitters and receptors is key for neural circuit analysis.

Purpose of the Study:

  • To demonstrate the functionality of novel cassettes for efficient recombineering of GAL4 and QF drivers.
  • To enable targeted gene expression control in specific neuronal populations using these recombineering methods.
  • To facilitate advancements in neural circuit mapping and manipulation techniques.

Main Methods:

  • Utilized recently available Drosophila genomic BAC libraries for recombineering large genes.

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  • Developed and tested cassettes for efficient recombineering of GAL4 and QF drivers.
  • Employed kanamycin selection for streamlined recombineering processes in Drosophila.

    • Main Results:

    • Successfully demonstrated the functionality of the developed cassettes for GAL4 and QF driver recombineering in Drosophila.
    • Showcased the feasibility of recombineering large genes, including those for neurotransmitters and receptors.
    • Established a method for precise genetic targeting in neural circuit research.

    Conclusions:

    • The developed cassettes provide an efficient and generalizable method for creating specific GAL4 and QF drivers.
    • This technique significantly advances the capability for neural circuit mapping and manipulation in Drosophila.
    • The cassettes hold potential for broader application in recombineering across different species.